Tomomitsu Tahara

Homozygous TRPV1 315C influences the susceptibility to functional dyspepsia.

Objectives Capsaicin/vanilloid (transient receptor potential vanilloid 1, (TRPV1) receptor has been shown to be expressed in gastrointestinal tract and play a role as a member of sensory ion channel superfamily. The G315C polymorphism affects the TRPV1 gene and alters its protein level. We aimed to investigate the effect of TRPV1 G315C polymorphism on functional dyspepsia (FD) in a Japanese population. Methods TRPV1 G315C polymorphism was genotyped in 98 subjects with no upper abdominal symptoms and 109 patients with FD. Severity of 7 upper gastrointestinal symptoms was assessed during cold water, and cold carbonated water drinking for randomly selected 20 healthy subjects. Results We found a significant inverse association between TRPV1 315CC genotype and FD [CC vs. others; odds ratio (OR)=0.40, 95% confidence interval (CI)=0.38-0.82]. We also found that the same genotype held a lower risk of both epigastric pain syndrome (OR=0.25, 95% CI=0.09-0.73), postprandial syndrome (OR=0.27, 95% CI=0.07-0.96) according to Rome III, and Helicobacter pylori positive FD (OR=0.28, 95% CI=0.10-0.79). The evolution of symptom severity scale of 7 total symptoms (P=0.004), and heavy feeling in stomach (P=0.02) during cold carbonated water drinking were significantly lower among 315CC genotypes compared with others. Conclusions Homozygous TRPV1 315C influences the susceptibility to FD through altering the upper gastrointestinal sensation.

Architecture of epigenetic reprogramming following Twist1-mediated epithelial-mesenchymal transition

BackgroundEpithelial-mesenchymal transition (EMT) is known to impart metastasis and stemness characteristics in breast cancer. To characterize the epigenetic reprogramming following Twist1-induced EMT, we characterized the epigenetic and transcriptome landscapes using whole-genome transcriptome analysis by RNA-seq, DNA methylation by digital restriction enzyme analysis of methylation (DREAM) and histone modifications by CHIP-seq of H3K4me3 and H3K27me3 in immortalized human mammary epithelial cells relative to cells induced to undergo EMT by Twist1.ResultsEMT is accompanied by focal hypermethylation and widespread global DNA hypomethylation, predominantly within transcriptionally repressed gene bodies. At the chromatin level, the number of gene promoters marked by H3K4me3 increases by more than one fifth; H3K27me3 undergoes dynamic genomic redistribution characterized by loss at half of gene promoters and overall reduction of peak size by almost half. This is paralleled by increased phosphorylation of EZH2 at serine 21. Among genes with highly altered mRNA expression, 23.1% switch between H3K4me3 and H3K27me3 marks, and those point to the master EMT targets and regulators CDH1, PDGFRα and ESRP1. Strikingly, Twist1 increases the number of bivalent genes by more than two fold. Inhibition of the H3K27 methyltransferases EZH2 and EZH1, which form part of the Polycomb repressive complex 2 (PRC2), blocks EMT and stemness properties.ConclusionsOur findings demonstrate that the EMT program requires epigenetic remodeling by the Polycomb and Trithorax complexes leading to increased cellular plasticity. This suggests that inhibiting epigenetic remodeling and thus decrease plasticity will prevent EMT, and the associated breast cancer metastasis.

DNA methylation as a molecular biomarker in gastric cancer

DNA methylation plays a significant role in gastric carcinogenesis. The CpG island methylator phenotype (CIMP) characterizes distinct subtypes of gastric cancer (GC) and the relationship between specific methylation patterns and clinicopathological features has been evaluated. Altered DNA methylation is also observed in Helicobacter pylori-infected gastric mucosa, and its potential utility for GC risk estimation has been suggested. The ability to detect small amounts of methylated DNA among tissues allows us to use DNA methylation as a molecular biomarker in GC in a variety of samples, including serum, plasma and gastric washes. The DNA methylation status of nontargeted tissue, particularly blood, has been associated with predisposition to GC. We focus on the recent development of DNA methylation-based biomarkers in GC.

Increased number of methylated CpG islands correlates with Helicobacter pylori infection, histological and serological severity of chronic gastritis.

Background Promoter hypermethylation of tumor suppressor genes is one of the major events in gastric carcinogenesis. Promoter hypermethylation is also present in non-neoplastic gastric epithelium as age-related phenomenon and some reports suggest the potential association between promoter hypermethylation and Helicobacter pylori infection. Here, we examined whether methylation of multiple promoter CpG islands would occur by H. pylori infection and correlate with histological or serological severity of chronic gastritis. Methods One hundred and ninety-one gastric mucosa samples were obtained by endoscopy. The promoter methylation status of the p14, p16, DAP-kinase and CDH1 genes were determined by methylation-specific-polymerase chain reaction. The degree of gastritis in the antrum was assessed according to the updated Sydney system in 150 participants. The pepsinogen (PG) I/II ratio was calculated based on the data of serum PG I and PG II levels measured by radioimmunoassay in 54 selected cases. Results CpG island methylation was found in 32.5% for p14, 35.1% for p16, 43.5% for DAP-kinase and 36.1% for CDH1, whereas non, 1, 2, 3, and all methylation of four promoter CpG sites were present in 46 (24.1%), 59 (30.9%), 46 (24.1%), 30 (15.7%), and 10 (5.2%) participants, respectively. A strong association between the increased number of methylated CpG islands and H. pylori infection was observed (P<0.0001). An increased number of methylated CpG islands was also associated with severity of neutrophil infiltration (P<0.0001), mononuclear cell infiltration (P<0.0001) and atrophy (P=0.0021) in all, and severity of neutrophil infiltration (P=0.0177) and mononuclear cell infiltration (P=0.0004) in H. pylori-positive participants. An increased number of methylated CpG islands correlated with lower PG I/II ratio in all (P=0.0105) and H. pylori-infected participants (P=0.074). Conclusion Multiple promoter CpG islands would be methylated by H. pylori infection, and an increased number of methylated CpG sites correlate with histological and serological severity of chronic gastritis.

Genetic polymorphisms of cyclooxygenase-1 (COX-1) are associated with functional dyspepsia in Japanese women.

BACKGROUND Prostaglandins (PGs), catalyzed from arachidonic acid by cyclooxygenase (COX), are involved in a variety of physiological processes in the stomach. COX-1 has been regarded as a constitutively expressed enzyme that generates prostaglandins for gastrointestinal integrity. We investigated the association between the potentially functional polymorphism T-1676C in the COX-1 gene promoter and functional dyspepsia (FD) in the Japanese population. METHODS The study was performed in 272 subjects (185 with no upper abdominal symptoms and 87 with FD). We employed the PCR-SSCP method to detect the gene polymorphism. RESULTS Overall, female sex had a 2-2.5-fold risk for the development of FD compared with male sex, whereas the COX-1 gene polymorphism and Helicobacter pylori infection were not associated with susceptibility to FD. However, in female subjects, -1676T allele carriers had a significantly increased risk for the development of FD (OR 2.70, 95%CI 1.04-6.99, p = 0.041), especially the epigastric pain syndrome (EPS) subgroup (OR 4.07, 95%CI 1.15-14.4, p = 0.029). CONCLUSIONS Our results provide the first evidence that the COX-1 gene polymorphism is significantly associated with the development of the EPS subgroup of FD in female subjects.

The usefulness of magnifying endoscopy with narrow-band imaging to distinguish carcinoma in flat elevated lesions in the stomach diagnosed as adenoma by using biopsy samples

BACKGROUND Therapeutic strategies for flat elevated (0-IIa) lesions in the stomach diagnosed as adenoma by biopsy are currently not established, because some difficulties have previously been reported in the evaluation of vascular patterns alone for the differential diagnosis between adenoma and carcinoma. OBJECTIVE We attempted to evaluate the 0-IIa lesions diagnosed as adenoma by using magnifying endoscopy with narrow-band imaging (MENBI) to distinguish them as either adenoma or carcinoma. SETTING Department of Gastroenterology, Fujita Health University. PATIENTS Fourteen adenomatous lesions (6 adenomas and 8 carcinomas confirmed postoperatively) diagnosed with preoperative biopsies from patients who had undergone endoscopic submucosal dissection were evaluated. INTERVENTIONS We selected 5 sites per lesion for MENBI. Selected sites were divided into superficial structures (SSs) and irregular microvascular patterns (IMVPs). MAIN OUTCOME MEASUREMENTS The rate of SSs and IMVPs in adenoma and carcinoma. RESULTS Significant SSs were tubular in the adenoma and unclear in the carcinoma. Regarding IMVP subcategories, (1) slight intrastructual irregular microvascular patterns (ISIMVPs) accounted for 97%, (2) severe ISIMVPs accounted for 0%, (3) fine networks accounted for 3%, and (4) corkscrews accounted for 0% of cases in the adenomas. The corresponding proportions in the carcinomas were (1) 40%, (2) 15%, (3) 45%, and (4) 0%. Severe ISIMVPs and fine networks were significant findings for carcinomas. LIMITATIONS The number of cases was limited. CONCLUSIONS Our combined evaluation method using MENBI offers the ability to establish proper therapeutic strategies for lesions that are difficult to identify as adenoma or carcinoma.

Multi-drug resistance 1 polymorphism is associated with reduced risk of gastric cancer in the Japanese population.

Background and Aims:  Host genetic factors play a key role in gastric carcinogenesis, but the mechanism has not been clarified. The multi‐drug resistance 1 (MDR1) gene mediates the expression of P‐glycoprotein, which has a role in active transport of various substrates, including xenobiotics, and thus has a protective function in various tissues and organs like gastrointestinal epithelial cells. C3435T polymorphism in exon 26 of the MDR1 gene influences P‐glycoprotein expression and activity in the gastrointestinal tract. We investigated the influences of MDR1 gene polymorphism on the risk of gastric cancer.

A genetic variant of the p22PHOX component of NADPH oxidase C242T is associated with reduced risk of functional dyspepsia in Helicobacter pylori-infected Japanese individuals.

Objectve Although inflammatory changes in the gastric mucosa are commonly observed in Japanese patients with functional dyspepsia (FD), detailed data regarding the relationship between the genetic regulatory factors of inflammation and FD are not available. Superoxide has been implicated in the pathogenesis of Helicobacter pylrori-related diseases through inflammation. Nicotinamide adenine dinucleotide phosphate oxidase, a major source of superoxide generation plays a critical role in H. pylori-related gastric inflammation. We aimed to clarify the association between C242T polymorphism of p22PHOX, an essential component of nicotinamide adenine dinucleotide phosphate oxidase and FD in a Japanese population. Methods Eighty-nine FD according to Rome III criteria and 95 asymptomatic participants enrolled in this study. The p22PHOX C242T polymorphism was determined by polymerase chain reaction restriction fragment length polymorphism. H. pylori infection status was examined by histology or antibody against H. pylori. Results Nonsignificant correlation was found between p22PHOX polymorphism and FD. In H. pylori positives, however, C242T carriers significantly associated lower risk of FD (25.9 vs. 6.2%; C/C vs. T carriers; odds ratio=0.19, 95% confidence interval=0.05–0.71, P=0.009). This significant association remained after logistic regression analysis with adjustment for sex and age (odds ratio=0.20, 95% confidence interval=0.05–0.73). No significant correlation was found between p22PHOX polymorphism and a different subgroup of FD. Conclusion Our data suggest that C242T carriers status is inversely related to the risk of FD in H. pylori-infected patients.

Methylome sequencing for fibrolamellar hepatocellular carcinoma depicts distinctive features

With the goal of studying epigenetic alterations in fibrolamellar hepatocellular carcinoma (FLC) and establish an associated DNA methylation signature, we analyzed LINE-1 methylation in a cohort of FLC and performed next-generation sequencing of DNA methylation in a training set of pure-FLCs and non-cirrhotic hepatocellular carcinomas (nc-HCC). DNA methylation was correlated with gene expression. Furthermore, we established and validated an epigenetic signature differentiating pure-FLC from other HCCs. LINE-1 methylation correlated with shorter recurrence-free survival and overall survival in resected pure-FLC patients. Unsupervised clustering using CG sites located in islands distinguished pure-FLC from nc-HCC. Major DNA methylation changes occurred outside promoters, mainly in gene bodies and intergenic regions located in the vicinity of liver developmental genes (i.e., SMARCA4 and RXRA). Partially methylated domains were more prone to DNA methylation changes. Furthermore, we identified several putative tumor suppressor genes (e.g., DLEU7) and oncogenes (e.g., DUSP4). While ∼70% of identified gene promoters gaining methylation were marked by bivalent histone marks (H3K4me3/H3K27me3) in embryonic stem cells, ∼70% of those losing methylation were marked by H3K4me3. Finally, we established a pure FLC DNA methylation signature and validated it in an independent dataset. Our analysis reveals a distinct epigenetic signature of pure FLC as compared to nc-HCC, with DNA methylation changes occurring in the vicinity of liver developmental genes. These data suggest new options for targeting FLC based on cancer epigenome aberrations.

Role of heat-shock protein (HSP) 70-2 genotype in peptic ulcer in Japanese population.

BACKGROUND/AIMS There have been reports showing the protective role of inducible heat-shock protein (HSP) 70 in gastric epithelial cells. An A to G transition at the 1267 position HSP70-2 gene has been shown to be associated with a different level of HSP70 mRNA expression. We aimed to clarify the effect of HSP70-2 polymorphism on the risk of peptic ulcer diseases in a Japanese population. METHODOLOGY A total of 519 subjects participated in this study. All subjects underwent upper gastroscopy. Restriction fragment length polymorphism analysis was performed for polymorphisms at 1267 of HSP70-2 gene in all the subjects. RESULTS After gastroscopy, 109, 53 and 357 subjects were diagnosed as gastric ulcer, duodenal ulcer and non-ulcer subjects, respectively. Although, there were no significant differences of HSP70-2 genotype distributions among nonulcer subjects, overall ulcer, gastric and duodenal ulcers when the subjects were divided into two groups according to age distribution, logistic regression analysis showed that the BB genotype increased the risk of duodenal ulcer in subjects 60 years and older. (Gender, status of H. pylori infection and NSAID use adjusted OR=3.12, 95%CI=1.33-7.35, p=0.009). CONCLUSIONS It appears that polymorphism of HSP70-2 gene is not directly associated with the susceptibility to peptic ulcer diseases but BB genotype is associated with an increased risk of duodenal ulcer in older subjects in the Japanese population.